biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

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Results 601 to 630 of 2229:

Amelioration of salt-induced oxidative stress in eggplant by application of 24-epibrassinolide

H. -D. Ding, X. -H. Zhu, Z. -W. Zhu, S. -J. Yang, D. -S. Zha, X. -X. Wu

Biologia plantarum 56:767-770, 2012 | DOI: 10.1007/s10535-012-0108-0

The effects of exogenous 24-epibrassinolide (EBR) on the growth, oxidative damage, antioxidant system and ion contents in eggplant (Solanum melongena L.) seedlings under salt stress were investigated. Eggplant seedlings were exposed to 90 mM NaCl with 0, 0.025, 0.05, 0.10 and 0.20 mg dm-3 EBR for 10 d. EBR, especially at concentration 0.05 mg dm-3, alleviated growth suppression caused by NaCl stress, decreased electrolyte leakage, superoxide production and content of malondialdehyde and H2O2 in NaCl-treated plants. EBR also increased activities of superoxide dismutase, guaiacol peroxidase, catalase and ascorbate peroxidase and the contents of ascorbic acid and reduced glutathione. Furthermore, we also found that Na+, Cl- contents were decreased, K+, Ca2+ contents and K+/Na+, Ca2+/Na+ ratios were increased in the presence of EBR under salt stress.

Genetic transformation of Citrus sinensis with Citrus tristeza virus (CTV) derived sequences and reaction of transgenic lines to CTV infection

F. R. Muniz, A. J. De Souza, L. C. L. Stipp, E. Schinor, W. Freitas Jr., R. Harakava, D. R. Stach-Machado, J. A. M. Rezende, F. A. A. Mourão Filho, B. M. J. Mendes

Biologia plantarum 56:162-166, 2012 | DOI: 10.1007/s10535-012-0035-0

Transgenic Citrus sinensis (L.) Osb. plants, cvs. Valencia and Hamlin, expressing Citrus tristeza virus (CTV) derived sequences were obtained by genetic transformation. The gene constructs were pCTV-CP containing the 25 kDa major capsid protein gene (CTV-CP), pCTV-dsCP containing the same CTV-CP gene in an intron-spliced hairpin construct, and pCTV-CS containing a 559 nt conserved region of the CTV genome. The transgenic lines were identified by PCR and the transgene integration was confirmed by Southern blot. Transgene mRNA could be detected in most transgenic lines containing pCTV-CP or pCTV-CS transgene. The mRNA of pCTV-dsCP transgene was almost undetectable, with very light bands in most analyzed plants. The transgene transcription appears to be closely linked to the type of gene construct. The virus challenge assays reveals that all transgenic lines were infected. However, it was possible to identify propagated clones of transgenic plants of both cultivars studied with a low virus titer, with values similar to the noninoculated plants (negative control). These results suggested that the transgenic plants present some level of resistance to virus replication. The higher number of clones with low virus titer and where mRNA could not be detected or was presented in a very light band was found for pCTV-dsCP-derived transgenic lines.

Do fern gametophytes have the capacity for irradiance acclimation?

B. Fernández-Marín, S. J. Arroyo Alfaro, J. M. Becerril, J. I. García-Plazaola

Biologia plantarum 56:351-356, 2012 | DOI: 10.1007/s10535-012-0097-z

Ferns present two alternant generations: sporophyte and gametophyte. In the present work we address the question of whether fern gametophytes have the potential to acclimate to different irradiances as vascular plants do. We studied the gametophytes of three different fern species belonging to the Aspleniaceae family with different ecological requirements (Asplenium trichomanes, Asplenium scoloprendrium and Ceterach officinarum). Fern spores were germinated and the gametophytes cultivated under photon flux density (PFD) of 10, 50 or 100 μmol m-2 s-1. From the early stages of spore germination (the formation of the 5-celled germinal filament), photosynthetic apparatus acclimates showing the typical patterns of photochemical responses to high or low PFD. In agreement with the photochemical pattern of acclimation, higher contents of xanthophyll cycle pigments and α-tocopherol was observed in plants grown under high PFD. The α/β-carotene ratio, used as indicator of the acclimation of the photosynthetic apparatus, also sustained the initial hypothesis except for A. trichomanes. We conclude that fern gametophytes display a complete array of photosynthetic and photoprotective traits that allow an effective acclimation to PFD.

Identification and characterization of a novel gene encoding myb-box binding zinc finger protein in Gossypium arboreum

M. Zahur, A. Maqbool, M. Irfan, A. Jamal, N. Shahid, B. Aftab, T. Husnain

Biologia plantarum 56:641-647, 2012 | DOI: 10.1007/s10535-012-0255-3

Zinc finger proteins are encoded by the genes chiefly involved in stress resistance hormone signal transduction of plants. In this study, a cDNA encoding a zinc finger transcription factor was isolated by the yeast one-hybrid system from Gossypium arboreum using the MYB-box element of the universal stress gene (GUSP1) promoter as bait. The corresponding protein (named GaZnF) can bind specifically to a 13 bp MYB-box region. The GaZnF cDNA is 1093 bp in length, including a 510 bp open reading frame. The predicted GaZnF protein contains ANI-A20 motifs and shares a high sequence similarity with zinc finger proteins from other plants. Spatial expression pattern of GaZnF was studied under drought, heavy metals and salt stresses through real-time PCR. The gene showed enhanced expression under each stress treatment with maximum transcript abundance in root tissues. The results support the hypothesis that G. arboreum zinc finger proteins are involved in plant response to drought, salt and heavy metal stresses.

Influence of UV-B radiation on young triticale plants with different wax cover

E. Skórska, W. Szwarc

Biologia plantarum 51:189-192, 2007 | DOI: 10.1007/s10535-007-0038-4

Ultraviolet-B radiation (biologically effective dose 2.6 kJ m-2 d-1) had negative influence on morphology and physiology of the young triticale plants. Plants exposed to UV-B were of lower height than control plants, their leaves were narrow, and the rate of net photosynthesis was decreased. The line RAH 336, which wax cover is lesser than that of traditional cultivar Magnat, was more susceptible to UV-B radiation, considering primary photosynthesis reactions, recorded by chlorophyll a fluorescence. An activation of protective mechanisms was observed: plants responded to UV-B by an increase of the content of UV-B absorbing compounds, and changes of antioxidant enzyme activities.

Transcriptome analysis of an invasive weed Mikania micrantha

Y. -L. Huang, X. -T. Fang, L. Lu, Y. -B. Yan, S. -F. Chen, L. Hu, C. -C. Zhu, X. -J. Ge, S. -H. Shi

Biologia plantarum 56:111-116, 2012 | DOI: 10.1007/s10535-012-0024-3

As an initial step towards understanding the molecular mechanisms by which plants become invasive, we present here the first transcriptome analysis for an invasive weed Mikania micrantha. The analysis was based on the 75-nucleotide short reads data generated by the Illumina Genome Analyzer II system. A total of 31 131 unique sequences were assembled de novo based on 8.7 million filtered paired-end sequence reads for the transcriptome of an individual M. micrantha growing in the field. 73 % of the unique sequences showed significant similarity to existing proteins in the NCBI database, and 21 448 could be grouped based on gene ontology assignments. Of particular interest are the sequences that share homology with genes involved in genome evolution, plasticity, secondary metabolism and defense responses.

Involvement of G6PDH in heat stress tolerance in the calli from Przewalskia tangutica and Nicotiana tabacum

H. Gong, G. Chen, F. Li, X. Wang, Y. Hu, Y. Bi

Biologia plantarum 56:422-430, 2012 | DOI: 10.1007/s10535-012-0072-8

Glucose-6-phosphate dehydrogenase (G6PDH) has been implicated in supplying reduced nicotine amide cofactors for biochemical reactions and in modulating the redox state of cells. In this study, the role of G6PDH in thermotolerance of the calli from Przewalskia tangutica and tobacco (Nicotiana tabacum L.) was investigated. Results showed that Przewalskia tangutica callus was more sensitive to heat stress than tobacco callus. The activity of G6PDH and antioxidant enzymes (ascorbate peroxidase, catalase, peroxidase and superoxide dismutase) in calli from Przewalskia tangutica and tobacco increased after 40 °C treatment, although two calli exhibited a difference in the degree and timing of response to heat stress. When G6PDH was partially inhibited by glucosamine pretreatment, the antioxidant enzyme activities and thermotolerance in both calli significantly decreased. Simultaneously, the heat-induced H2O2 content and the plasma membrane NADPH oxidase activity were also reduced. Application of H2O2 increased the activity of G6PDH and antioxidant enzymes in both calli. Diphenylene iodonium, a NADPH oxidase inhibitor, counteracted heatinduced H2O2 accumulation and reduced the heat-induced activity of G6PDH and antioxidant enzymes. Moreover, exogenous H2O2 was effective in restoring the activity of G6PDH and antioxidant enzymes after glucosamine pretreatment. Western blot analysis showed that G6PDH gene expression in both calli was also stimulated by heat and H2O2, and blocked by DPI and glucosamine under heat stress. Taken together, under heat stress G6PDH promoted H2O2 accumulation via NADPH oxidase and the elevated H2O2 was involved in regulating the activity of antioxidant enzymes, which in turn facilitate to maintain the steady-state H2O2 level and protect plants from the oxidative damage.

Visualisation of xylem sap flow direction in isolated fine lateral roots and estimation of the xylem sap osmotic potential

M. Fritz, R. Ehwald

Biologia plantarum 56:749-756, 2012 | DOI: 10.1007/s10535-012-0121-3

Xylem sap outflow from fine lateral roots (FLRs) isolated from hydroponically grown young maize (Zea mays L.) plants was visualized by local brightening of test solutions contrasted with purified Indian ink particles. Flow into the vessels was indicated by the adsorption of Evans Blue in their walls. The fraction of the FLRs able to exude xylem sap in a mineral medium with 30 mM mannitol decreased with increasing incubation time. This change was strongly retarded, when the FLRs were incubated in a medium containing glucose instead of mannitol. There was a broad range of variation of the osmotic potential of the test solutions (Ψso), wherein the fraction of the FLRs showing an initially reversed flow of the xylem sap varied between zero and unity. A median (M) of the osmotic potential of the xylem sap in FLRs (Ψsx) was estimated. It represents the value of Ψso that was lower than Ψsx in half of the roots of a sample before their transfer to the test solutions (Ψsxo). M was dependent on the osmotic potential of the medium used for growth or pre-incubation of the FLRs. Its value was not dependent on the molecular size of the osmolytes used to adjust Ψso, including dextran 8, which is excluded from cell walls. In all of the studied plants, M was lower than the osmotic potential of the xylem sap collected from the root before isolation of the FLRs. To explain this finding it is assumed that FLRs with Ψsxo > M had a higher hydraulic conductivity and a larger volume contributed to the exuded sap than those with Ψsx < M.

Physiological and ultrastructural effects of lead on tobacco

R. Alkhatib, J. Maruthavanan, S. Ghoshroy, R. Steiner, T. Sterling, R. Creamer

Biologia plantarum 56:711-716, 2012 | DOI: 10.1007/s10535-012-0241-9

The effects of lead toxicity on leaf gas exchange, chlorophyll content, chlorophyll fluorescence, chloroplast ultrastructure, and opening of stomata were examined in tobacco (Nicotiana tabacum L.) plants. Plants were grown in a growth chamber for 7 d in Hoagland nutrient solution supplemented with 0.0 (control), 5, 10, 25, 50, 100, 300 and 500 μM Pb(NO3)2. Plants treated with 5, 10, and 25 μM Pb(NO3)2 exhibited no abnormalities. Root and shoot growth, net photosynthetic rate and stomatal conductance were significantly reduced in plants treated with 100, 300 and 500 μM Pb(NO3)2. In plants treated with 500 μM Pb(NO3)2, the majority of stomata were closed. The effect of Pb(NO3)2 on chlorophyll content and chlorophyll fluorescence parameters was negligible. However, in plants exposed to 100, 300, and 500 μM Pb(NO3)2, the mesophyll cells showed altered chloroplasts with disrupted thylakoid membranes.

Exogenous 4-hydroxybenzoic acid and salicylic acid modulate the effect of short-term drought and freezing stress on wheat plants

E. Horváth, M. Pál, G. Szalai, E. Páldi, T. Janda

Biologia plantarum 51:480-487, 2007 | DOI: 10.1007/s10535-007-0101-1

Exogenous salicylic acid has been shown to confer tolerance against biotic and abiotic stresses. In the present work the ability of its analogue, 4-hydroxybenzoic acid to increase abiotic stress tolerance was demonstrated: it improved the drought tolerance of the winter wheat (Triticum aestivum L.) cv. Cheyenne and the freezing tolerance of the spring wheat cv. Chinese Spring. Salicylic acid, however, reduced the freezing tolerance of Cheyenne and the drought tolerance of Chinese Spring, in spite of an increase in the guaiacol peroxidase and ascorbate peroxidase activity. The induction of cross tolerance between drought and freezing stress was observed: drought acclimation increased the freezing tolerance of Cheyenne plants and cold acclimation enhanced the drought tolerance. The induction of drought tolerance in Cheyenne was correlated with an increase in catalase activity.

Isolation and functional characterization of MxCS1: a gene encoding a citrate synthase in Malus xiaojinensis

D.-G. Han, Y. Wang, L. Zhang, L. Ma, X.-Z. Zhang, X.-F. Xu, Z.-H. Han

Biologia plantarum 56:50-56, 2012 | DOI: 10.1007/s10535-012-0015-4

Iron is one of the essential micronutrients required by all living organisms. In this study, we isolated a gene encoding putative citrate synthase (CS) from Malus xiaojinensis, designated as MxCS1. The MxCS1 gene encodes a protein of 473 amino acid residues with a predicted molecular mass of 52.5 kDa and a theoretical isoelectric point of 8.67. The expression of MxCS1 was enriched in the leaf rather than in phloem and root, however, its expression was hardly detected in the xylem. The gene expression was strongly induced by Fe stress treatment in the M. xiaojinensis seedlings. Over-expression of MxCS1 improved Fe deficiency tolerance in transgenic Arabidopsis. We argued that MxCS1 is a new member of the CS genes, and it may function as a regulator in response to iron stress in plants.

Isolation and expression of a cold-responsive gene PtCBF in Poncirus trifoliata and isolation of citrus CBF promoters

L. G. He, H. L. Wang, D. C. Liu, Y. J. Zhao, M. Xu, M. Zhu, G. Q. Wei, Z. H. Sun

Biologia plantarum 56:484-492, 2012 | DOI: 10.1007/s10535-012-0059-5

C-repeat/dehydration-responsive element binding factor (CBF) plays important roles in cold response network in plants. Here, one member of CBF coding gene family in trifoliate orange (Poncirus trifoliata), designated as PtCBF, was isolated. Semi-quantitative reverse transcription-polymerase chain reactions showed up-regulation of PtCBF not only under low temperature but also induced by abscisic acid. Additionally, the CBF genomic fragments in four citrus species including trifoliate orange, sweet orange (Citrus sinensis), pummel (Citrus grandis) and rough lemon (Citrus jambhiri) were isolated with complete open reading frames. According to the results of alignment analysis between full length cDNA and genomic DNA sequences in trifoliate orange, there were no introns in PtCBF. Moreover, the results of multiple sequence alignment analysis and phylogenetic analysis on putative protein sequences suggested that the AP2 DNA binding domains and CBF signature sequences were highly conserved in four citrus CBF proteins. Finally, the CBF promoters in above citrus species were isolated, which provides some information concerning promoter function.

Cadmium modulates NADPH oxidase activity and expression in sunflower leaves

M. D. Groppa, M. P. Ianuzzo, E. P. Rosales, S. C. Vázquez, M. P. Benavides

Biologia plantarum 56:167-171, 2012 | DOI: 10.1007/s10535-012-0036-z

The production of reactive oxygen species (ROS) and the ways by which ROS are generated are very important facts related to heavy metal toxicity in plants. In this work, superoxide anion (O2 .-) generation diminished in cadmium treated sunflower (Helianthus annuus L.) leaf discs, and this reduction was time and Cd-concentration dependent. In line with these findings, we observed that NADPH-dependent oxidase activity was significantly inhibited by 0.1 and 0.5 mM Cd2+ treatments and the expression of the NADPH oxidase putative gene related to O2 .- synthesis in sunflower leaves was 83 % inhibited by 0.1 mM CdCl2 and almost completely depleted by 0.5 mM CdCl2.

Micropropagation of Pinus peuce

D. Stojičić, D. Janoąević, B. Uzelac, V. Čokeąa, S. Budimir

Biologia plantarum 56:362-364, 2012 | DOI: 10.1007/s10535-012-0099-x

In Pinus peuce zygotic embryo culture grown on Gresshoff and Doy (1972; GD) basal medium, 2.22 μM benzyladenine (BA) was superior in promoting adventitious bud induction during 4 weeks comparing to kinetin or BA + kinetin. Shoot elongation was achieved on half-strength GD medium devoid of plant growth regulators and containing activated charcoal. Pulse treatment with 1 mM indole-3-butyric acid (IBA) for 2 h, followed by transfer to half-strength GD medium, produced the most efficient rooting. Rooted shoots were transplanted to the greenhouse and plantlets continued to grow and developed into phenotypically normal plants. Up to 10 plants per explant can be obtained within 36 weeks from culture initiation.

Salt- and osmotic stress-induced choline monooxygenase expression in Kochia scoparia is ABA-independent

E. B. Kalinina, B. K. Keith, A. J. Kern, W. E. Dyer

Biologia plantarum 56:699-704, 2012 | DOI: 10.1007/s10535-012-0132-0

Choline monooxygenase (CMO) is the first regulatory enzyme in the biosynthetic pathway for glycine betaine, an effective osmoprotectant in Kochia scoparia, a highly drought- and salt-tolerant species. In seedlings, CMO transcript levels are rapidly increased in response to both NaCl and osmotic stress treatments. The mRNA level in shoots was substantially higher than in roots. The rapid induction seen in whole plants was in contrast to the apparent down-regulation observed in suspension-cultured K. scoparia cells in response to the same salt stress. Treatment with exogenous abscisic acid (ABA) or fluridone shows that CMO induction proceeds via an ABA-independent signal transduction pathway. Examination of the CMO upstream regulatory region reveals a number of stress response-related elements, some of which may be involved in the stress tolerance shown by this species.

Transformation with a gene for myo-inositol O-methyltransferase enhances the cold tolerance of Arabidopsis thaliana

B. Zhu, R. -H. Peng, A. -S. Xiong, J. Xu, X. -Y. Fu, W. Zhao, X. -F. Jin, X. -R. Meng, J. -J. Gao, R. Cai, Q. -H. Yao

Biologia plantarum 56:135-139, 2012 | DOI: 10.1007/s10535-012-0029-y

In this study, we report a function of myo-inositol-O-methyltransferase (Imt1) in response to low temperature stress using transgenic Arabidopsis thaliana. Imt1 gene was constructed identical to the Imt1 gene from a halophyte Mesembryanthemum crystallinum. After cold stress, the Imt1 transgenic plants exhibited stronger growth than the wild type plants. The elevated cold tolerance of the Imt1 over-expressing plants was confirmed by the lower electrolyte leakage and accumulation of malondialdehyde, but higher proline and soluble sugar contents in transgenic than wild type plants.

Ascorbate deficient semi-dwarf asfL1 mutant of Lathyrus sativus exhibits alterations in antioxidant defense

D. Talukdar

Biologia plantarum 56:675-682, 2012 | DOI: 10.1007/s10535-012-0245-5

An ascorbate-deficient semi-dwarf mutant asfL-1 was detected in 250 Gy γ-ray treated grass pea (Lathyrus sativus L.) cv. BioR-231. The mutant contained only 42 % of leaf and 20 % of root ascorbate content of mother control (MC). I investigated the possible causes of ascorbate deficiency and its effect on growth and antioxidant defense in control and 150 mM NaCl-treated seedling after 60 d growth period. Ascorbate deficiency was due to significant reduction in activities of monodehydroascorbate reductase and dehydroascorbate reductase as well as increase in ascorbate oxidase, leading to considerable decrease in redox state. Despite low ascorbate pool and decrease in ascorbate peroxidase activity, shoot and root biomass production in asfL-1 mutant were similar to MC plants, even at NaCl treatment. High accumulation of glutathione (GSH) coupled with high activities of GSH reductase, catalase, GSH peroxidase and peroxidase in both tissues of the mutant permitted efficient recycling of GSH and scavenging of H2O2 through well integrated catalase/peroxidase system, despite high superoxide dismutase activity under NaCl treatment. The collapse of this system led to inhibition of growth in NaCl-treated mother plants. Together, the results suggested that asfL-1 plants undertook a major reshuffle in its antioxidant defense machinery, which effectively counterbalanced the negative impact of ascorbate deficiency and remained unperturbed by NaCl treatment to maintain normal growth and biomass production.

Fatty acid content during reconstitution of the photosynthetic apparatus in the air-dried leaves of Xerophyta scabrida after rehydration

K. Georgieva, A. Ivanova, S. Doncheva, S. Petkova, D. Stefanov, E. Péli, Z. Tuba

Biologia plantarum 55:581-585, 2011 | DOI: 10.1007/s10535-011-0130-7

Desiccation of Xerophyta scabrida caused considerable damage of chloroplast ultrastructure together with a complete loss of chlorophyll. Upon rehydration, the relative water content of the pale-green leaves almost reached that of the dark-green ones, however, the Chl content and photosynthetic activity remained lower. The process of reconstitution of the photosynthetic apparatus in the re-greening leaves was accompanied by changes in fatty acid (FA) content. The amount of the FA methyl esters was more than 2-fold higher in the green leaves as compared to the dry ones and slightly increased after rehydration in the pale-green leaves. Among the three main fatty acids in the leaves, oleic, palmitic and linoleic acid, the latter increased more than 3-fold during rehydration. This acid is concentrated mainly in the glycolipids and this was an indirect indication for the restoration of the photosynthetic apparatus. Our results showed that rehydration of X. scabrida led to a decrease of the saturated FA in parallel with an increase of the unsaturated FA, thus indicating increased membrane permeability. The observed changes in the lipid content can be considered as a characteristic feature of X. scabrida and most probably of other poikilochlorophyllous species.

Nitric oxide is involved in the regulation of ascorbate and glutathione metabolism in Agropyron cristatum leaves under water stress

C. Shan, F. He, G. Xu, R. Han, Z. Liang

Biologia plantarum 56:187-191, 2012 | DOI: 10.1007/s10535-012-0040-3

This study investigated the regulation of ascorbate and glutathione metabolism by nitric oxide in Agropyron cristatum leaves under water stress. The activities of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), L-galactono-1,4-lactone dehydrogenase (GalLDH) and γ-glutamylcysteine synthetase (γ-ECS), and the contents of NO, reduced ascorbic acid (AsA), reduced glutathione (GSH), total ascorbate and total glutathione increased under water stress. These increases were suppressed by pretreatments with NO synthesis inhibitors N G-nitro-L-arginine methyl ester (L-NAME) and 4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). However, application of L-NAME and cPTIO to plants sufficiently supplied with water did not affect the activities of above mentioned enzymes and the contents of NO and above mentioned antioxidants. Pretreatments with L-NAME and cPTIO increased the malondialdehyde (MDA) content and electrolyte leakage of plants under water stress. Our results suggested that water stress-induced NO is a signal that leads to the upregulation of ascorbate and glutathione metabolism and has important role for acquisition of water stress tolerance.

Mitochondrial minisatellite polymorphisms in fodder and sugar beets reveal genetic bottlenecks associated with domestication

Y. Yoshida, M. Matsunaga, D. Cheng, D. Xu, Y. Honma, T. Mikami, T. Kubo

Biologia plantarum 56:369-372, 2012 | DOI: 10.1007/s10535-012-0101-7

Historically, sugar beets were selected from fodder beets. We used mitochondrial minisatellite loci to analyze cytoplasmic genetic diversity in fodder beet and sugar beet. Among the 8 sugar beet accessions examined we identified 3 multi-locus haplotypes. These 3 haplotypes were a subset of 5 haplotypes identified among the 29 fodder beet accessions examined. All but one haplotype in fodder beet comprised, in turn, a subset of 12 haplotypes identified previously in leaf beets. Such apparent decreases in cytoplasmic genetic diversity must result from genetic bottlenecks associated with domestication and the ensuing breeding processes. We also detected the haplotype associated with the male-sterile Owen cytoplasm of sugar beet in the fodder beet gene pool. Furthermore, the presence of a 39 kDa protein associated with the Owen cytoplasm was confirmed in two fodder beet plants by Western blot analysis. These results lead us to speculate that the Owen cytoplasm may have originated in fodder beet, from which sugar beet was derived.

Molecular cloning and characterization of nucleoside diphosphate kinase 1 cDNA in tea

G. R. Prabu, K. Thirugnanasambantham, A. K. A. Mandal, A. Saravanan

Biologia plantarum 56:140-144, 2012 | DOI: 10.1007/s10535-012-0030-5

Nucleoside diphosphate kinase (NDPK) operates in the homeostasis of cellular nucleoside triphosphate (NTP) pools and the cytosolic NDPK1 is the main NDPK isoform in plants, accounting for more than 70 % of total NDPK activity in plant. A full length cDNA (697 bp), designated as CsNDPK1 was cloned from Camellia sinensis (L.) O. Kuntze leaves. Sequence analysis of CsNDPK1 shows several motifs, binding and catalytic sites which are highly conserved among other NDPKs. Southern blot analysis revealed that tea genome has two copies of CsNDPK1. Transcription pattern analysis indicated that CsNDPK1 is expressed in all tissues examined, but expressed more in buds than in other organs.

Frequencies and variation in cytosine methylation patterns in diploid and tetraploid cytotypes of Paspalum notatum

M. P. Rodriguez, G. D. L. Cervigni, C. L. Quarin, J. P. A. Ortiz

Biologia plantarum 56:276-282, 2012 | DOI: 10.1007/s10535-012-0087-1

Paspalum notatum Flügge is a grass species organized as an agamic complex. The objective of the current research was to survey the frequencies and variation of cytosine methylation at CCGG sequences in diploid and tetraploid genotypes, and to determine the occurrence of methylation changes associated with tetraploidization by using methylation-sensitive amplification polymorphism (MSAP) markers. No differences were found in the average proportions of methylated CCGG sites between cytotypes, but methylation patterns were significantly more variable in tetraploids. In both groups of plants, epigenetic and non-epigenetic variation correlated significantly when compared by Mantel tests. The evaluation of 159 common MSAP markers showed that 18.86 % of them differed in their methylation status in the different ploidies. Dendrogram analysis, reflecting epigenetic distances, showed that the four diploids and one experimentally-obtained sexually-reproducing tetraploid, grouped together. MSAP analysis performed on a diploid plant and its autotetraploid derivative showed that new epialleles emerged after tetraploidization. Sequencing of several MASP markers showed homologies with low copy genes, non-coding sequences and transposon/retrotransposon elements.

Characterization of a drought inducible trithorax-like H3K4 methyltransferase from barley

D. Papaefthimiou, A. S. Tsaftaris

Biologia plantarum 56:683-692, 2012 | DOI: 10.1007/s10535-012-0125-z

Histone H3 lysine 4 methylations catalyzed by histone lysine methyltransferases (HKMTs), like the Arabidopsis thaliana ATX1 and ATX2, are important epigenetic modifications related to chromatin decondensation and gene activation. In order to study this epigenetic mechanism in monocot cereal plants, we performed homology searches of ATX1 and ATX2 against the Brachypodium distachyon L. Beauv and rice (Oryza sativa L. spp. japonica) genomes, discovering single homologues for each cereal crop representing both Arabidopsis sequences. Using this information, we employed the rolling circle amplification - rapid amplification of cDNA ends (RCA-RACE) method to isolate, clone and characterize HvTX1 from RNA extracted from barley (Hordeum vulgare L.) tissues and studied its expression during seed development and under drought stress. The cloned cDNA sequence contained a 3 093 bp ORF homologous to ATX1 and ATX2. Characterization of the translated HvTX1 transcript sequence revealed the multi-domain nature of the putative protein, including all conserved regions characteristic for ATX1 and ATX2. By comparative genomic analysis and homology searches in EST databases we located, with high probability, the gene coding for HvTX1 on the barley chromosome 5H. Constant elevation of HvTX1 expression was observed during seed development. Expression of HvTX1 after drought stress was analyzed by quantitative real-time polymerase chain reaction (qPCR) in two different barley cultivars with varying drought stress tolerance, revealing HvTX1 drought-induction in a tolerance-specific manner.

Phylogenetic, structure and expression analysis of ABC1Ps gene family in rice

S. G. Yang, T. Li, M. Liu, S. J. Gao, S. Y. Yang, L. Li, C. W. Yang

Biologia plantarum 56:667-674, 2012 | DOI: 10.1007/s10535-012-0247-3

The ABC1 protein family (ABC1P), a new family of putative kinases, widely existed in procaryote and eucaryote. A comprehensive genome-wide analysis was carried out in this study to find all ABC1Ps in rice (Oryza sativa subsp. japonica). We identified 15 ABC1P genes in rice. All the ABC1Ps contained an ABC1 domain of about 120 conserved amino acid residues and conserved kinase motifs-VAIK (VAVK, VAMK) and DFG (DEG). The phylogenetic analysis showed that all the ABC1Ps were grouped into three subgroups, and formed a total of 12 sister pairs. Conserved motifs analysed by MEME program indicated that almost all ABC1Ps contains motifs 1, 3, 7, 8 and 9. Predictably, the ABC1Ps were localized in mitochondria or chloroplasts, which implied that the ABC1Ps might be involved in energy metabolism in plants. RT-PCR assays demonstrated that all 15 ABC1P genes were active, and some of them were affected by abiotic stresses (NaCl, high temperature, methyl viologen, abscisic acid and cadmium).

Improving low-temperature tolerance in sugarcane by expressing the ipt gene under a cold inducible promoter

N. G. Belintani, J. T. S. Guerzoni, R. M. P. Moreira, L. G. E. Vieira

Biologia plantarum 56:71-77, 2012 | DOI: 10.1007/s10535-012-0018-1

Sugarcane is cultivated in tropical and subtropical regions where cold stress is not very common, but lower yields and reduced industrial quality of the plants are observed when it occurs. In our efforts to enhance cold tolerance in sugarcane, the gene encoding the enzyme isopentenyltransferase (ipt) under control of the cold inducible gene promoter AtCOR15a was transferred via biolistic transformation into sugarcane (Saccharum spp.) cv. RB855536. Semiquantitative RT-PCR using GAPDH encoding glyceraldehyde-3-phosphate dehydrogenase as the normalizer gene showed the increased expression of the ipt gene under cold stress. The detached leaves of genetically modified plants subjected to low temperatures showed visible reduction of leaf senescence in comparison to non-transgenic control plants. Induced overexpression of ipt gene also enhanced cold tolerance of non-acclimated whole plants. After being subjected to freezing temperature, leaf total chlorophyll contents of transgenic plants were up to 31 % higher than in wild type plants. Also, lower malondialdehyde content and electrolyte leakage indicated less damage induced by cold in transgenic plants. Thus, the expression of ipt driven by the stress inducible COR15a promoter did not affect plant growth while providing a greater tolerance to cold stress.

The role of non-symbiotic haemoglobin and nitric oxide homeostasis in waterlogging tolerance in Vigna species

R. K. Sairam, K. Dharmar, V. Chinnusamy, S. Lekshmy, R. Joshi, P. Bhattacharya

Biologia plantarum 56:528-536, 2012 | DOI: 10.1007/s10535-012-0064-8

The objective of this study was to examine the role of nitrate reductase, nitric oxide and non-symbiotic hemoglobin in imparting waterlogging tolerance in mung bean genotypes. Experiment was conducted with two cultivated mung bean [Vigna radiata (L.) Wilczek] genotypes T 44 (tolerant) and Pusa Baisakhi (susceptible) and a highly tolerant wild species Vigna luteola (Jacq.) Benth. The content of nitric oxide increased up to 6 d of waterlogging in Vigna luteola and T 44, and up to 4 d of treatment in Pusa Baisakhi. Increase in nitrate reductase (NR) activity was observed only up to 4 d of waterlogging in Vigna luteola and T 44, and up to 2 d of treatment in Pusa Baisakhi, and thereafter the activity decreased in all the genotypes. The increase in NO content and NR activity was greater in Vigna luteola and T 44 than in Pusa Baisakhi. Non-symbiotic hemoglobin (NSHb) and cNR mRNA expressions were observed only in waterlogging treated roots of Vigna luteola and T 44, while very little expression was observed in control plants of Vigna luteola and T 44, and in control and waterlogged plants of Pusa Baisakhi. PCR bands of Hb and cNR were cloned, and nucleotide and deduced amino acid sequences were obtained and conserved regions and domains were identified using database.

Isolation and molecular characterization of cinnamate 4-hydroxylase from apricot and plum

A. Pina, T. Zhebentyayeva, P. Errea, A. Abbott

Biologia plantarum 56:441-450, 2012 | DOI: 10.1007/s10535-012-0114-2

Cinnamate 4-hydroxylase (C4H) is the second enzyme in the phenylpropanoid pathway which participates in the synthesis of numerous phenylpropanoid compounds such as flavonoids, lignins, suberins and others. We identified a gene putatively coding for Class I C4H in apricot and plum and we analyzed the expression pattern of this gene under different apricot/plum graft combinations with different degree of compatibility. The full-length cDNA is 1 739 bp with a 1 515 bp open reading frame encoding a protein of 504 amino acids. Like other C4Hs, the predicted C4H polypeptides included conserved domains of cytochrome P450. The genomic sequence of the apricot C4H gene was interrupted by two introns 335 bp and 904 bp long. Several regulatory motifs including P-, A-, L- and H-boxes, which were conserved across phenylpropanoid metabolism-related genes in higher plants, were found in a 1 300 bp upstream promoter region of the apricot C4H gene. A phylogenetic analysis showed that all Prunus sequences clustered together and were closely related to Malus and Rubus C4H genes. The transcription of Class I PruC4H was detected in all the examined graft combinations, which suggested its rather constitutive character.

Interspecific somatic hybrids between Solanum bulbocastanum and S. tuberosum and their haploidization for potato breeding

M. Iovene, R. Aversano, S. Savarese, I. Caruso, A. Di Matteo, T. Cardi, L. Frusciante, D. Carputo

Biologia plantarum 56:1-8, 2012 | DOI: 10.1007/s10535-012-0008-3

Protoplast fusion between incongruent Solanum bulbocastanum and S. tuberosum haploids was accomplished to produce hybrids combining elite traits from both parents. We identified 11 somatic hybrids out of 42 regenerants analyzed through ISSR markers. Some hybrids had loss or gain of fragments compared to the parents, likely due to rearrangements and deletions of chromosome segments after fusion, and/or to somaclonal variation during hybrid regeneration. Increased heterotic vigor for some traits as well as high diversity was observed as the effect of both ploidy and fusion combination. Microsporogenesis analysis indicated the occurrence of multivalent configurations and several meiotic abnormalities, such as chromosomes bridges and various spindle orientations. Since all hybrids were sterile, in vitro anther culture was employed for haploidization as a possible strategy to overcome barriers to hybridizations. Haploids were obtained from all the tetraploid S. bulbocastanum (+) S. tuberosum somatic hybrids tested, although with differences in both the number of embryos per 100 anthers cultured and the number of differentiated green plantlets. This is the first report on the successful production of haploid plants from S. bulbocastanum (+) S. tuberosum hybrids.

Effect of 24-epibrassinolide on drought stress-induced changes in Chorispora bungeana

Y. H. Li, Y. J. Liu, X. L. Xu, M. Jin, L. Z. An, H. Zhang

Biologia plantarum 56:192-196, 2012 | DOI: 10.1007/s10535-012-0041-2

Brassinosteroids (BRs) have been proposed to increase the resistance of plants to drought stress. The effect of foliar application of 0.1 μM 24-epibrassinolide (EBR) on chlorophyll (Chl) content, photosystem 2 (PS 2) photochemistry, membrane permeability, lipid peroxidation, relative water content (RWC), proline content, and the antioxidant system in drought-stressed Chorispora bungeana plants was investigated. The results showed that polyethylene glycol (PEG) induced water stress decreased RWC, Chl content and variable to maximum Chl fluorescence ratio (Fv/Fm) less in plants pretreated with EBR than in non-pretreated plants. In addition, lipid peroxidation, measured in terms of malondialdehyde content, membrane permeability and proline content in drought-stressed plants were less increased in EBR pretreated plants, while antioxidative enzyme activities and reduced ascorbate and glutathione contents were more increased in EBR pretreated than in non-pretreated plants. These results suggested that EBR could improve plant growth under drought stress

Na+/H+ antiport activity in plasma membrane and tonoplast vesicles isolated from NaCl-treated cucumber roots

K. Kabała, M. Janicka-Russak

Biologia plantarum 56:377-382, 2012 | DOI: 10.1007/s10535-012-0103-5

Sodium/proton antiporter activity in the plasma membrane and tonoplast of cucumber seedling roots treated with 200 mM NaCl for 24 h was determined. It was observed that plasma membrane and tonoplast antiporter activity was only present in membranes from salt-treated plants. In addition, the plasma membrane antiporter protein was present in membranes after induction with NaCl, whereas tonoplast antiporter protein was observed in control and at elevated level in NaCl-treated plants. Moreover, based on the affinity of studied antiporter proteins to sodium ions, it could be assumed that excess sodium ions are firstly translocated from the cytosol to the vacuole and then excluded to the apoplast through the plasma membrane.

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